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为了满足转基因玉米成分检测中阳性对照不易获得及不易长久保存的缺陷,通过分析公共数据库资源,建立了包含玉米内标准基因z SSIIb及遗传转化中常用的Ca MV35S启动子、nos启动子、Ca MV35S终止子、nos终止子的的标准分子。灵敏度检测发现,在检测体系中含有1 000个分子时,可以非常清楚扩增到所有5个片段的目的条带。随机挑选我国批准进口的7种转基因玉米,对标准分子进行验证,结果表明,在相应的阳性样品和标准分子中都能扩增到特异的目的条带,构建的标准分子可以满足作为转基因玉米筛选检测中阳性对照的要求。
In order to meet the deficiencies of the positive control not easy to be obtained and not easy to be stored for long time in the detection of genetically modified corn, we established a genetic linkage map that contains the standard gene z SSIIb in maize and CaMV35S promoter, nos promoter, Ca MV35S Terminator, nos terminator standard molecule. Sensitivity tests revealed that when the assay contained 1 000 molecules, the band of interest amplified to all 5 fragments was very clear. Seven kinds of transgenic maize approved by our country were selected at random to validate the standard molecules. The results showed that specific target bands could be amplified in the corresponding positive samples and standard molecules, and the constructed standard molecules could meet the requirement of screening for transgenic corn Test positive control requirements.